LOCUS V037231 6864 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V037231 VERSION V037231 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 235..614 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 615..818 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 863..881 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 927..1667 /label="HES4(NM_001142467)" /note="HES4(NM_001142467)" /gene="HES4" CDS 1707..2426 /codon_start=1 /note="mammalian codon-optimized" /product="the original enhanced GFP (Yang et al., 1996)" /transl_table=1 /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV LLEFVTAAGITLGMDELYK*" /label="EGFP" polyA_signal 2464..2688 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2734..3162 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3176..3505 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3572..4366 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" polyA_signal 4540..4661 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4710..4726) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4734..4750 /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG)." /label="lac operator" promoter complement(4758..4788) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4803..4824 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5112..5697) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5868..6728) /codon_start=1 /gene="bla" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /product="β-lactamase" /transl_table=1 /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA SLIKHW*" /label="AmpR" promoter complement(6729..6833) /gene="bla" /label="AmpR promoter" ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gtttaaagcc actgggatcc gccaccatgg ccgcagacac gccggggaaa ccgagcgcct 961 cgccgatggc aggagcgccg gccagcgcca gccggacccc agacaagccc cggagcgcgg 1021 ccgagcaccg caaggtgggg tcccggccgg gcgtgagggg ggcgaccggg gggcgggagg 1081 gacgcgggac tcagccggtg cccgacccgc agtcctccaa gccggtcatg gagaagcggc 1141 gccgagcgcg tattaacgag agcctcgctc agctcaaaac cctcatcctg gacgccctca 1201 gaaaagagag ctcccgccac tcgaagctgg agaaggcgga catcctggag atgaccgtga 1261 gacacctgcg gagcctgcgt cgcgtgcagg tgacggccgc gctcagcgcc gaccccgccg 1321 ttctgggcaa gtaccgcgcc ggcttccacg agtgtctggc ggaggtgaac cgcttcctgg 1381 ccggctgcga gggcgtcccg gccgacgtgc gctcccgcct gctgggccac ctggcagcct 1441 gcctgcgcca gctgggaccc tcccgccgcc cggcctcgct gtccccggct gcccccgcag 1501 aggccccagc gcccgaggtc tacgcgggcc gcccgctgct gccatcgctc ggcggcccct 1561 tccctctgct cgcgccgccg ctgctgccgg gtctgacccg ggcgctgccc gccgccccca 1621 gggcggggcc gcagggcccg ggtgggccct ggaggccgtg gctgcgcatc tcgagtggcg 1681 gtggaggaag cggcggaggt ggatctatgg tgagcaaggg cgaggagctg ttcaccgggg 1741 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg 1801 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg 1861 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct 1921 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag 1981 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg 2041 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca 2101 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct 2161 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca 2221 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg 2281 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc 2341 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc 2401 tcggcatgga cgagctgtac aagtagtcta gagggcccgt ttaaacccgc tgatcagcct 2461 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga 2521 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt 2581 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg 2641 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg 2701 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg 2761 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg 2821 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc 2881 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa 2941 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc 3001 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac 3061 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt 3121 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg 3181 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca 3241 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat 3301 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc 3361 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat 3421 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt 3481 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc 3541 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg 3601 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg 3661 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa 3721 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct 3781 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga 3841 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc 3901 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac 3961 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc 4021 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact 4081 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga 4141 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg 4201 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga 4261 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga 4321 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg 4381 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc 4441 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc 4501 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat 4561 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg 4621 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg 4681 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat 4741 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc 4801 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga 4861 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt 4921 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg 4981 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac 5041 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg 5101 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca 5161 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc 5221 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc 5281 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag 5341 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc 5401 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca 5461 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg 5521 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg 5581 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct 5641 ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa 5701 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg 5761 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga 5821 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta 5881 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc 5941 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg 6001 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga 6061 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt 6121 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt 6181 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc 6241 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc 6301 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca 6361 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag 6421 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg 6481 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa 6541 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa 6601 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga 6661 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga 6721 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg 6781 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt 6841 ccccgaaaag tgccacctga cgtc //